輔酶A（CoA）檢測(cè)試劑盒
產(chǎn)品名稱：輔酶A（CoA）檢測(cè)試劑盒
英文名稱：Coenzyme A （CoA） Assay Kit
貨號(hào)：K367-100
品牌：原裝進(jìn)口
包裝：100次分析
保存建議: 請(qǐng)?jiān)诮邮盏皆?/font>輔酶A（CoA）檢測(cè)試劑盒產(chǎn)品后，置于-20℃保存.
產(chǎn)品背景: 輔酶A是由半胱氨酸、泛酸和ATP組成的小單元構(gòu)成的.輔酶A是很多生物過(guò)程的重要分子，它在脂肪酸的合成和氧化降解、檸檬酸循環(huán)過(guò)程中丙酮酸的氧化等生物過(guò)程中都起重要作用.輔酶A的一個(gè)主要功能就是攜帶酰基基團(tuán)，而所攜帶的酰基基團(tuán)中，最重要的是乙酰基團(tuán).攜帶了乙?；妮o酶A稱為乙酰輔酶A.這個(gè)乙酰基團(tuán)將會(huì)被用于合成各種生物大分子，比如膽固醇、乙酰膽堿、褪黑素以及TCA循環(huán)中的許多中間產(chǎn)物.
產(chǎn)品描述: 輔酶A檢測(cè)試劑盒提供了一種檢測(cè)生物樣品中CoA水平的方案，操作便捷，結(jié)果精確.檢測(cè)實(shí)驗(yàn)中，樣本內(nèi)游離的輔酶A被轉(zhuǎn)變成一種中間產(chǎn)物，該中間產(chǎn)物可以與OxiRed探針發(fā)生反應(yīng)，最終的反應(yīng)產(chǎn)物顯現(xiàn)顏色（λ = 570 nm）和熒光（Ex=535/Em=587 nm）.本輔酶A檢測(cè)試劑盒可檢測(cè)的輔酶A范圍是0.1-10 nmol（2.5-250 M的濃度范圍）.
產(chǎn)品特點(diǎn) 1、步驟簡(jiǎn)單，只需要1小時(shí)左右即可完成檢測(cè)；2、操作快速并且方便； 3、該輔酶A檢測(cè)分析試劑盒靈敏、穩(wěn)定，也適用于高通量分析
Histone deacetylases (HDACs) represent a large family of enzymes identified as key
regulators of nucleosomal histone acetylation, a major event that controls eukaryotic gene
transcription and are classified into three groups. They are believed to be involved in important
biological activities, such as cell differentiation, proliferation, apoptosis, and senescence. In
BioVision’s HDAC8 Inhibitor screening Kit, HDAC8 deacetylates the substrate (R-H-K(Ac)-K(Ac)-
AFC) followed by an enzymatic cleavage to release the AFC molecule, which can be detected
fluorometrically (Ex/Em = 380/500 nm). In the presence of an HDAC8 inhibitor, the deacetylation
will be impeded which prevents the cleavage of the substrate to release AFC. The kit provides a
rapid, simple, sensitive, and reliable test, which is also suitable for high throughput screening of
HDAC8 inhibitors. Trichostatin A is included as a control inhibitor to compare the efficacy of the test
inhibitors.
II. Kit Contents:
III. Storage and Handling:
Remove the vial of HDAC8 and store the kit at -20°C, protected from light. Allow HDAC8 Assay
Buffer to warm to room temperature before use. Briefly centrifuge vials before opening. Read the
entire protocol before performing the assay.
HDAC-8 Enzyme: Store enzyme at -80°C. Use within 2 months.
HDAC-8 Substrate: Upon thawing aliquot and store at -20°C, use within 2 months.
Developer and Trichostatin A: Store at -20°C; use within 2 months.
HDAC-8 Assay Buffer: Store at 4°C.
IV. HDAC-8 Inhibitor Screening Assay Protocol:
1. Enzyme preparation: For each well, prepare a total 50 μl HDAC-8 Enzyme Solution:
48 μl HDAC8 Assay Buffer
2 μl HDAC8 Enzyme
2. Screen compounds, inhibitor control and blank control preparations:
Dissolve candidate inhibitors into a proper solvent. Dilute to 4X the desired test concentration with
HDAC8 Assay Buffer. For Trichostain A Inhibitor Control, dilute 1:9 with HDAC8 Assay Buffer. Add
25 μl of diluted test inhibitors, Trichostatin A or HDAC8 Assay Buffer into HDAC8 enzyme wells as
sample screen, Inhibitor Control (Trichostatin A), or Enzyme Control (No inhibitor). Mix well, and
incubate for 10 minutes at 37°C.
3. Background well: Add 75 μl HDAC8 Assay Buffer into a blank well.
4. Substrate preparation: For each well, prepare a total 25 μl Substrate Solution:
23 μl Assay Buffer
2 μl Substrate
Mix, add 25 μl of the substrate to all sample wells. Mix, incubate 60 min at 37°C.
5. Developer: Add 10 μl into each well (including background well); Mix well. Incubate for 5 min
at 37°C.
6. Measurement: Read Ex/Em=380/500 nm as RBG, RS, and REC for background well, sample
wells or inhibitor control and enzyme control wells, respectively.
7. Calculation: The RFU of fluorescence generated by hydrolyzation of substrate is ?RFU of
samples (or inhibitor control) ?RFU S = RS – RBG, ?RFU of Enzyme Control = ?RFU EC = REC
– RBG. Set the ?RFU EC as the 100%, and calculate the relative % Inhibition of the test
inhibitors as:
% Inhibition =
?RFU EC? ?RFU S
?RFU_EC
× 100%
RELATED PRODUCTS:
HDAC1 thru HDAC11 Antibodies HDAC Family Antibody Set
DiscoveryPak? HDAC Inhibitor Set HDAC Activity Assay Colorimetric Kit
HDAC Activity Assay Fluorometric Kit HAT Activity Assay Colorimetric Kit
HAT (P/CAF) Active Human Recombinant HAT-1 thru HAT-3 Antibodies
Trichostatin A HDAC8 Activity Assay Kit
HDAC8, human recombinant HDAC3 Activity Assay Kit
HDAC3 Inhibitor Screening Assay Kit HDAC3, human recombinant
FOR RESEARCH USE ONLY! Not to be used on humans.
0
50
100
0 100 200
% Relative Activity
TSA (nM)
TSA Inhibition of HDAC-8 Activity
Components 100 assays Cap Color Part Number
HDAC-8 Assay Buffer 25 ml WM K368-100-1
HDAC-8 Substrate 200 μl Red K368-100-2
HDAC-8 Enzyme 200 μl Green K368-100-3
Developer 1 ml Orange K368-100-4
Trichostatin A (100 μM) 200 μl Blue K368-100-5
如與英文說(shuō)明書有異，以英文說(shuō)明書為準(zhǔn)。
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